Part:BBa_K3394000
Coding Sequence of Endo5a cellulase (for E. coli)
This part consists oF the coding region of Endo5A, an endo-1,4-beta-glucanase. Endo5a breaks down cellulose by cleaving the internal 1-4 beta glucose linkages in the cellulose chain.
The complete cellulose hydrolysis reaction involves this enzyme along with several other enzymes: endo-1,4-beta-glucanases, exo-1,4-beta-glucanases (or cellobiohydrolases) and beta-glucosidases.
Endo5a is a relatively small protein with an optimal operating temperature of 50o C and an optimal pH range of 6.0-7.0. The NCBI GenBank number for Endo5a is HQ657203.1 (DNA sequence), AEB00655.1 (amino acid sequence).
We learned about the cellulase from a past iGEM team – the British Columbia 2016 team which fused it to the surface layer of Caulobacter crescentus (BBa_K2139001). In our project, we have performed codon optimization for its expression in Escherichia coli, and the sequence was synthesized by IDT for us.
Endo5A was isolated from the bacterial flora found in the gut of a cotton bollworm (Helicoverpa armigera) which secretes a variety of plant-hydrolyzing enzymes.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 898
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 898
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 898
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 898
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 898
- 1000COMPATIBLE WITH RFC[1000]
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